RAD Tag Sequencing of Nigronia serricornis (Say) (Megaloptera: Corydalidae), On and Off Reservoirs Emily M. Berezowski Faculty Sponsor: Jeffrey S. Heilveil, Ph.D. Biology Department, SUNY College at Oneonta, Oneonta, NY

ABSTRACT Reservoirs are a great asset to humans, but can have several adverse effects on surrounding wildlife. Changes in a landscape as a result of reservoir creation may cause changes in interactions between organisms and ultimately, change dispersal and gene flow, which can isolate populations. This research will use an aquatic fishfly, Nigronia serricornis (Say) (Megaloptera: Corydalidae), to determine if reservoir creation altered gene flow between populations. Restriction-site Associated DNA (RAD) sequencing will be used to sequence the DNA of individuals from equidistant sets of populations on tributaries flowing into the reservoirs and ones flowing directly into the Delaware, unaffected by reservoirs. Differences in gene flow patterns may indicate an anthropogenic effect on local ecosystems. If gene flow patterns for N. serricornis have changed, other species that utilize these waterways may also be affected.

INTRODUCTION SIGNIFICANCE OF RESEARCH

The creation of reservoirs alters landscapes Change in gene flow and population which can have negative effects on local population connectivity in N. serricornis could suggest changes interactions. The saw-combed fishfly, Nigronia in other species that utilize these waterways, and serricornis (Say) (Megaloptera: Corydalidae), may be a that information could be of use in the planning of species that has been affected by reservoir creation, conservation projects or other environmental as it is stream dwelling for most of its life. planning. A previous student project used microsatellite loci developed at SUNY Oneonta to determine the effect of the Pepacton Reservoir on populations of Nigronia serricornis, and found that the reservoir played a role in genetic isolation. This research is continuing the aforementioned study at sites along the Pepacton Reservoir and the Ashokan Reservoir, using Restriction Site Associated DNA Sequencing (sensu Peterson et al. 2012), in the place of microsatellite data. After mapping out partial Fig. 2: Pepacton Reservoir, Delaware County, NY. genomes, the data will be used to examine variation in the populations. If reservoirs are affecting these populations there should be a reduction in gene flow between sites on each of the two large reservoirs. METHODS

Sixteen Nigronia serricornis (Say) were © E. Berezowski collected at sites on tributaries flowing into the Fig. 3: Nigronia serricornis (Say) Pepacton Reservoir (See Fig. 1). Genomic DNA was extracted from prothoracic tissue. A Double Digest reaction was performed as per Peterson LITERATURE CITED et al. (2012), using ECOR1 and MSP1 as Davey, John W, and Mark L Blaxter. "Radseq: Next-Generation Population Genetics." Briefings In Functional Genomics 9.5-6 (2010): 416-423. MEDLINE with Full Text. Web. 9 Oct. 2014. restriction enzymes. Each individual was Peterson, Brant K., Jesse N. Weber, Emily H. Kay, Heidi S. Fisher, and Hopi E. Hoekstra. "Double Digest RADseq: An assigned a unique four base-pair long adapter Inexpensive Method for De Novo SNP Discovery and Genotyping in Model and Non-Model Species." PLoS ONE 7.5 (2012): n. pag. sequence for identification after RAD Stevenson, Rachel Marie, and Jeffrey S. Heilveil. "Characterization Of Eight Polymorphic Microsatellite Markers For The Sequencing, attached individually by ligation. Saw-Combed Fishfly, Nigronia Serricornis (Say) (Megaloptera: Corydalidae)." Conservation Genetics Resources 4 (2013): 1111. Academic OneFile. Web. 6 Oct. 2014. Samples were purified, pooled, and size selected to fragments of 300-800 base pair long ACKNOWLEDGEMENTS sequences by gel extraction. The sample was PCR amplified and prepared for RAD We would like to thank J. Salvino for site location sequencing at Cornell Genomics Laboratory. information, and N. Pedisich, J. Pennings J. Piacente, and A. Rothfuss for assistance in the field. This Fig. 1: Map of specific sites. Inset shows location of reservoir in NYS. research is funded by a grant from the SUNY College Senate Committee on Research.